Kaempferia parviflora Extracellular Vesicle Loaded with Clarithromycin for the Treatment of Helicobacter pylori Infection

Int J Nanomedicine. 2024 Feb 27:19:1967-1983. doi: 10.2147/IJN.S444686. eCollection 2024.

Abstract

Purpose: Kaempferia parviflora extracellular vesicles (KPEVs) have been reported as promising nanovesicles for drug delivery. This study aimed to load clarithromycin (CLA) into KPEVs (KPEVS-CLA) and determine the physical properties, drug-releasing efficiency, gastric cell uptake, anti-H. pylori activities, and anti-inflammatory responses in comparison with free CLA and KPEVs.

Methods: The size and surface charge of KPEVs-CLA were evaluated using dynamic light scattering and visualized using a transmission electron microscope. The encapsulation efficiency (EE%), loading capacity (LC%), and drug release of KPEVs-CLA were examined using HPLC. Anti-H. pylori growth and anti-adhesion were evaluated. IL-8 gene expression, NF-κB signaling proteins, and anti-inflammatory profiles were examined using qRT-PCR, Western blotting, and Bio-Plex immunoassay, respectively. Anti-chemotaxis was then examined using a Transwell assay.

Results: KPEVs-CLA were intact and showed a negative surface charge similar to that of KPEVs. However, slightly enlarged KPEVs were observed. CLA was successfully loaded into KPEVs with EE of 93.45% ± 2.43%, LC of 9.3% ± 3.02%. CLA release in the PBS and gastric mimic buffer with Fickian diffusion (n ≤ 0.43) according to Korsmeyer-Peppas kinetic model (R2=0.98). KPEVs-CLA was localized in the gastric cells' cytoplasm and perinuclear region. Anti-H. pylori growth and anti-H. pylori adhesion of KPEVs-CLA were compared with those of free CLA with no cytotoxicity to adenocarcinoma gastric cells. KPEVs-CLA significantly reduced IL-8, G-CSF, MIP-1α, and MIP-1β levels. Moreover, KPEVs-CLA showed a superior effect over CLA in reducing G-CSF, MIP-1α, and NF-κB phosphorylation and monocyte chemotactic activities.

Conclusion: KPEVs serve as potential carriers of CLA. They exhibited a higher efficiency in inhibiting gastric cell inflammation mediated by H. pylori infection than free CLA. The establishment of KPEVs-CLA as a nanodrug delivery model for H. pylori treatment could be applied to other plant extracellular vesicles or loaded with other cancer drugs for gastric cancer treatment.

Keywords: Helicobacter pylori; Kaempferia parviflora; clarithromycin; extracellular vesicle; inflammation.

MeSH terms

  • Anti-Inflammatory Agents / pharmacology
  • Antibodies
  • Chemokine CCL3
  • Clarithromycin
  • Granulocyte Colony-Stimulating Factor
  • Helicobacter Infections* / drug therapy
  • Helicobacter pylori*
  • Humans
  • Interleukin-8
  • NF-kappa B

Substances

  • Clarithromycin
  • Chemokine CCL3
  • Interleukin-8
  • NF-kappa B
  • Antibodies
  • Anti-Inflammatory Agents
  • Granulocyte Colony-Stimulating Factor

Grants and funding

This work was supported by the Thailand Research Fund through the Royal Golden Jubilee Ph.D. program (Grant No. PHD/0217/2559), 90th Anniversary Chulalongkorn University Fund (Ratchadaphiseksomphot Endowment Fund, GCUGR1125633043D) and Thailand Science Research and Innovation (TSRI) (CU_FRB640001_01_37_1). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.